Small-molecule sensing and capture by de novo protein receptors followed by fluorescence spectroscopy and mass spectrometry.

通过从头蛋白质受体进行小分子传感和捕获，然后进行荧光光谱和质谱分析。

• 批准号: 2293551
• 金额: --
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2293551
• 关键词: Small; molecule; sensing; capture; de

Proteins already have an incredibly versatile array of structures and functions in nature and our understanding of them has enabled the design of new structures. De novo protein design has enabled the creation of proteins for use in industrial biotechnologies or engineering, free of the selective pressures found in nature. One protein fold, coiled coils, are one such example of successful de novo design targets. Their structure can be described parametrically which has allowed for computational design, and the discovery of new oligomeric states as of yet unseen in nature. Higher order coiled coils possess a continuous pore running through their structure, named a-helical barrels (aHBs). These aHBs have been shown to be incredibly thermostable and tolerable of several mutations in the lumen orientated residues. aHBs have already been exploited to perform simple chemical modifications, full catalytic cycles and the capturing of small-molecules. The latter of which is only just beginning to be explored. At the current understanding, lipids are the most attractive targets for aHBs. The lipidome can be used to identify many diseases through the use of characteristic biomarkers including cancers, obesity, Alzheimer's disease and diabetes. Treated lipids samples are often analysed through the use of mass spectroscopy coupled with a chromatographic separation method. However, such metabolomic studies can be intensive, requiring complex instrumentation with a large perquisite of knowledge to analyse the results. Here, we aim to identify specific aHB:ligand binding partners in complex fluids in order to produce simple to read, quantitative colorimetric diagnostic tests through the use of aHBs arrays in tandem with environment-sensitive dyes. aHBs by their nature attract hydrophobic compounds with some specificity on compound size. Enhancing our understanding of how the general binding occurs would further enable peptide design with unique binding-partner types in mind. The differential sensing of particular macromolecules can enable the fingerprinting of different conditions and the presence, or lack thereof, of unique compounds. The result would be a hyper-stable, mutable and easily readable diagnostic array with applications in industry and medicine.

蛋白质在自然界中已经具有令人难以置信的多样性的结构和功能，我们对它们的理解使新结构的设计成为可能。从头开始的蛋白质设计使得能够创造用于工业生物技术或工程的蛋白质，而不受自然界中发现的选择性压力的影响。一个蛋白质折叠，卷曲，是成功从头设计目标的一个这样的例子。它们的结构可以用参数描述，这使得计算设计和发现新的低聚状态成为可能，而这些状态在自然界中还是未知的。高阶螺旋盘管具有贯穿其结构的连续气孔，称为a-螺旋管(AHBs)。这些aHBs已被证明具有令人难以置信的热稳定性，并且对管腔导向残基的几个突变具有耐受性。AHBs已经被利用来执行简单的化学修饰、完整的催化循环和捕获小分子。后者才刚刚开始探索。根据目前的理解，脂类是aHBs最有吸引力的靶点。通过使用包括癌症、肥胖症、阿尔茨海默氏症和糖尿病在内的特征生物标记物，脂体可以用来识别许多疾病。经过处理的脂类样品通常通过使用质谱仪和层析分离方法进行分析。然而，这样的代谢组研究可能是密集的，需要复杂的仪器和大量的知识来分析结果。在这里，我们的目标是识别复杂流体中特定的ahb：配体结合伙伴，以便通过将aHBs阵列与环境敏感染料结合使用来产生简单易读的定量比色诊断测试。AHBs本质上吸引疏水化合物，在化合物大小上具有一定的特异性。加强我们对一般结合如何发生的理解将进一步使肽设计考虑到独特的结合伙伴类型。对特定大分子的差示传感可以对不同条件和独特化合物的存在或缺乏进行指纹识别。其结果将是一种超稳定、易变和易读的诊断阵列，在工业和医学上有应用。